1. APh161 - Lecture 15: Molecular Motors Rob Phillips California Institute of Technology

2. Mitotic Spindle Organization Cilia and Flagella Assembly and Dynamics Formation of Golgi and ER Networks Vesicle Transport Molecular Motors

3. Rogues Gallery of Motor Action: Rotary Motors Show Berg movie Show Yasuda et al. movie

4. Rogues Gallery of Motor Action: Translocation

5. Rogues Gallery of Motor Action: Translational Motor 1

6. Rogues Gallery of Motor Action: Translational Motor 2 - Muscles See Hugh Huxley review on website

7. Rogues Gallery of Motor Action: Translational Motor 2 - Muscles Heuser lab – Washington University

8. Organellar Transport (Hirokawa, Science 1998)

9. Rogues Gallery of Motor Action: Translational Motor 2

10. Myosin V Speed: 350 nm/s ATPase: 5.0 1/s Kinesin Speed: 850 nm/s ATPase: 44.0 1/s Dynein Speed: -1250 nm/s ATPase: 2.0 1/s **ALL INVOLVED IN VESICLE TRANSPORT** Translational Motors

11. How We Know: Gliding Motility Assays Show Vale movie

12. Dynamics of Molecular Motors Science, Vol 300, Issue 5628, 2061-2065, 27 June 2003 - Yildiz et al. Show Gelles, Selvin movie

13. Stepping Kinetics Spudich et al., PNAS 2000

14. Kinesin Data Fig. 3. Kinesin velocity as a function of [ATP] under external loads, F, fixed by a force clamp. The plots, from the top down, are for F = 0, 1.05, 3.59, 4.60, and 5.63 pN, respectively. Data from Block and colleagues (9): solid curves, N = 2 fits; dashed curves, N = 2 predictions (see text) (Fisher and Kolomeisky, PNAS)

15. Kinesin Data continued Fig. 4. Fits to the data of Block and colleagues (9) (and predictions) for velocity as a function of load for fixed concentrations of ATP. Note the inflection points at low [ATP] and convex profile at saturating [ATP]. (Fisher and Kolomeisky, PNAS)

16. Kinesin Randomness Data Fig. 5. Randomness data from Block and colleagues (9) and theoretical fits (A) as a function of external load, F, at fixed [ATP] (note that the two data points at F 5.7 pN and [ATP] = 2 mM appear separately in Block and coworkers: see figure 4 a and b of ref. 9, respectively) and (B) as a function of [ATP] at fixed loads of, from top down, F = 5.69 pN ( ), 5.35 and 4.60 pN (dashed-line predictions), 3.59 pN ( ), and 1.05 pN ( ).

17. Motors and States

18. What do we mean by the states?

19. Translocation Motors

20. Measurements on Rate of DNA Ejection DNA injection from T5 into vesicle Injection rate in λ (Baldeschweiler et al.) (Bohm et al.)

21. Measurements on Rate of DNA Ejection Phage Hypothesized Mechanism Genome Length (kbp) Ejection time (sec) Av. Ejection rate (kbp/sec) lambdaPressure48.5600.8 T4Pressure169305.6 T7Enzyme406000.06 T5 Pressure+ Enzyme 1213600.3 phi29Pressure+ Enzyme 1918000.05

22. Calculating the Ejection Time The mean first passage time is

23. The Time of Ejection receptor vesicle No protein interactions